myd88 antibody - bsa free  (Bio-Techne corporation)

Journal: International Journal of Molecular Sciences

Article Title: Cilastatin Attenuates Acute Kidney Injury and Reduces Mortality in a Rat Model of Sepsis

doi: 10.3390/ijms26167927

Figure Lengend Snippet: Effects of cilastatin on cecal ligation and puncture (CLP)-induced Toll-like receptor 4 (TLR4)/Myeloid differentiation factor 88 (MyD88) pathway activation. ( A ) Western blot of TLR4 in the renal cortex and ( B ) respective quantification corrected by the loading control. Results are expressed as mean ± SEM, n = 4–5 animals per group, * p ≤ 0.0094 vs. all groups. ( C ) Renal mRNA expression of TLR4 . Results are expressed as mean ± SEM, n = 4–5 animals per group, * p ≤ 0.046 vs. all groups. ( D ) Immunolocalization of TLR4 through immunofluorescence staining (green) in kidney sections (magnification 20× bar 50 µm). Cilastatin significantly decreased the TLR4 levels that were increased by CLP (arrowheads). ( E ) Serum measurement of LPS Binding Protein (LBP). Results are expressed as mean ± SEM, n = 4 animals per group, * p ≤ 0.0074 vs. all groups. ( F ) Western blot of MyD88 in renal cortex and ( G ) densitometric analysis corrected by the loading control. Results are expressed as mean ± SEM, n = 4–5 animals per group, * p ≤ 0.0094 vs. all groups. ( H ) Renal mRNA expression of MyD88 . Results are expressed as mean ± SEM, n = 5–7 animals per group. ( I ) Immunolocalization (upper image) and immunofluorescence staining (lower image) of MyD88 in renal tissue (magnification 20× bar 50 µm) and ( J ) semi-quantification. Results are expressed as mean ± SEM, n = 6 animals per group, * p ≤ 0.0017 vs. all groups. Note increased the staining in CLP rats (arrowhead and asterisks) and the change in the pattern of that staining compared with CLP + cilastatin and control groups. Cilastatin significantly decreased the MyD88 levels that were increased by CLP, although no significant differences in RNA levels were found. The data were analyzed using ANOVA. Cil: cilastatin; a.u., arbitrary units.

Article Snippet: Sections were then incubated overnight at 4 °C in a humidified chamber with primary antibodies diluted in PBS-T, 4% bovine serum albumin, and 1% host serum in which the secondary antibody was obtained as follows: mouse anti-RelA/NF-κB p65 monoclonal antibody (Novus Biologicals, Minneapolis, MN, USA [dilution 1:100], ref. NB100-56712); mouse anti-TLR4 (25) monoclonal antibody (Santa Cruz Biotechnology [dilution 1:50], ref. sc-293072); and rabbit anti-MyD88 polyclonal antibody (Novus Biologicals [dilution 1:50], ref. NB100-56698).

Techniques: Ligation, Activation Assay, Western Blot, Control, Expressing, Immunofluorescence, Staining, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Cilastatin Attenuates Acute Kidney Injury and Reduces Mortality in a Rat Model of Sepsis

doi: 10.3390/ijms26167927

Figure Lengend Snippet: Summary image of the effects of cilastatin on the Toll-like receptor 4 (TLR4)/Myeloid differentiation factor 88 (MyD88)/Nuclear Factor-κB/NF-κB) pathway and nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing protein 3 (NLRP3) inflammasome in cecal ligation and puncture (CLP)-induced inflammation and acute kidney injury. The TLR4 signaling complex is organized and localized in the cholesterol lipid rafts on the brush border apical side of renal proximal tubular epithelial cells, geographically close to the renal dehydrodipeptidase I (DHP-I) enzyme. In the left panel, with cholesterol rafts intact, damage-associated molecular patterns (DAMPS) or pathogen-associated molecular patterns (PAMPS), such as bacterial lipopolysaccharide (LPS), activate the TLR4/MyD 88 pathway, leading to the activation of NF-κB and pro-inflammatory cytokine and chemokine production. NF-κB is also involved in the activation of the NLRP3 inflammasome that leads to the activation of caspase-1, which, in turn, regulates the processing of inflammatory pro-cytokines to active cytokines (such as interleukin (IL)-1β), amplifying inflammatory damage and cell death, which exacerbates kidney injury. In the right panel, cilastatin binding to the membrane DHP-I in cholesterol lipid rafts causes modifications in the rafts, preventing the correct assembly and activation of the TLR4 complex and reducing the activation of NF-κB and the NLRP3 inflammasome. Thus, the renal cell is protected. K+, potassium; Ca2+, calcium; TIRAP, TIR Domain Containing Adaptor, Protein; IκB, inhibitor of nuclear factor kappa B; TNFα, tumor necrosis factor alpha; GSDMD, Gasdermin D.

Article Snippet: Sections were then incubated overnight at 4 °C in a humidified chamber with primary antibodies diluted in PBS-T, 4% bovine serum albumin, and 1% host serum in which the secondary antibody was obtained as follows: mouse anti-RelA/NF-κB p65 monoclonal antibody (Novus Biologicals, Minneapolis, MN, USA [dilution 1:100], ref. NB100-56712); mouse anti-TLR4 (25) monoclonal antibody (Santa Cruz Biotechnology [dilution 1:50], ref. sc-293072); and rabbit anti-MyD88 polyclonal antibody (Novus Biologicals [dilution 1:50], ref. NB100-56698).

Techniques: Binding Assay, Ligation, Activation Assay, Membrane

Journal: Antioxidants (Basel, Switzerland)

Article Title: Antioxidant and Anti-Apoptotic Neuroprotective Effects of Cinnamon in Imiquimod-Induced Lupus.

doi: 10.3390/antiox13070880

Figure Lengend Snippet: Figure 1. (A): Schematic representation of the experimental design, showing imiquimod treatment and cinnamon administration for the different groups: sham, sham treated with cinnamon (Sham Cinna), imiquimod-induced lupus (Lupus), imiquimod-induced lupus with cinnamon treatment (Lupus Cinna), and imiquimod-induced lupus with cinnamon administration starting 2 weeks before induction and continued after induction (Cinna Lupus Cinna). (B) Hippocampal immunofluorescence (scale bars 15 µm, magnification ×100) with quantification of IgG accumulation in the different groups. (C,D) Antibody array (C) with heatmap (D) of oxidative stress markers, TLR7 and MYD88, for the different groups. (E–K) Quantification of p-NRF/NRF, p-FOXO3/FOXO3, TLR7/β-actin, MYD88/β- actin, p-NOS3/NOS3, SOD1/β-actin, and SOD2/β-actin. (L,M) Western blot and quantification of p-FOXO3/FOXO3. (N,O) Western blot and quantification of p-NRF/NRF. a.u.: arbitrary units. * p < 0.05 and ** p < 0.01.

Article Snippet: The additional primary antibodies were: anti-Toll-like receptor 7 TLR7 (ab24184; Abcam, Cambridge, UK), anti-Caspase 3 (ab13847; Abcam, Cambridge, UK), anti-Caspase 8 (ab108333; Abcam, Cambridge, UK), anti-BID (ab62469; Abcam, Cambridge, UK), t-BID (ab10640; Abcam, Cambridge, UK), MyD88 (NB100-56698SS) (Novus Biologicals Bio-Techne, Minneapolis, MN, USA), NOS3 (sc-654), 8-OHDG (sc-66036), SOD1 (sc-101523), SOD2 (sc133134) (Santa Cruz Biotechnology, Dallas, TX, USA), and NOS3 (Ser1177) (#9571) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Immunofluorescence, Ab Array, Western Blot

Journal: Antioxidants (Basel, Switzerland)

Article Title: Antioxidant and Anti-Apoptotic Neuroprotective Effects of Cinnamon in Imiquimod-Induced Lupus.

doi: 10.3390/antiox13070880

Figure Lengend Snippet: Figure 4. (A) Schematic representation of in vitro treatment of cultured hippocampal cells with imiquimod, imiquimod with cinnamon, or the plasma of different mice groups. (B–J) Antibody arrays, heatmap, and quantification of TLR7, MYD88, and oxidative stress markers in hippocampal cells treated with the plasma of different mice groups. (K–S) Antibody arrays, heatmap, and histograms of TLR7, MYD88, and oxidative stress markers in cultured hippocampal cells treated with imiquimod, cinnamon with imiquimod, or sham. a.u.: arbitrary units. * p < 0.05, ** p < 0.01.

Article Snippet: The additional primary antibodies were: anti-Toll-like receptor 7 TLR7 (ab24184; Abcam, Cambridge, UK), anti-Caspase 3 (ab13847; Abcam, Cambridge, UK), anti-Caspase 8 (ab108333; Abcam, Cambridge, UK), anti-BID (ab62469; Abcam, Cambridge, UK), t-BID (ab10640; Abcam, Cambridge, UK), MyD88 (NB100-56698SS) (Novus Biologicals Bio-Techne, Minneapolis, MN, USA), NOS3 (sc-654), 8-OHDG (sc-66036), SOD1 (sc-101523), SOD2 (sc133134) (Santa Cruz Biotechnology, Dallas, TX, USA), and NOS3 (Ser1177) (#9571) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: In Vitro, Cell Culture, Clinical Proteomics

Journal: Journal of ethnopharmacology

Article Title: Effect of Echinacea purpurea (L.) Moench and its extracts on the immunization outcome of avian influenza vaccine in broilers.

doi: 10.1016/j.jep.2023.117306

Figure Lengend Snippet: Fig. 4. EP and EE affects the immune gene expression in jejunum: a) relative TLR4 mRNA expression, b) relative MyD88 mRNA expression, c) relative TPAF6 mRNA expression, d) relative AP-1 mRNA expression. All data were presented as mean ± SD(n = 6). *p < 0.05,**p < 0.01,***p < 0,001 compared to the group C; #p < 0.05,##p < 0.01,###p < 0,001 compared to the group V and intergroup.

Article Snippet: Name Company Cat. no. Dilution TLR4 Rabbit pAb Bioss bs-20379R 1:1000 MyD88 Rabbit pAb Novus Biologicals NB100-56698 1:1000 ERK1/2Rabbit pAb Abmart T55487 1:1000 JNK1/2/3Rabbit pAb Abmart T40073 1:1000 TRAF6Rabbit pAb ABclonal A16991 1:1000 GAPDHRabbit pAb ABclonal AC001 1:9000 Goat anti-Rabbit IgG Zenbio H&L 1:5000 X. Wang et al. Journal of Ethnopharmacology 319 (2024) 117306 On day 7 after vaccination, the FCR of the VE and VEE groups was significantly lower compared to the groups C and V (P < 0.05).

Techniques: Gene Expression, Expressing

Journal: Journal of ethnopharmacology

Article Title: Effect of Echinacea purpurea (L.) Moench and its extracts on the immunization outcome of avian influenza vaccine in broilers.

doi: 10.1016/j.jep.2023.117306

Figure Lengend Snippet: Fig. 5. Effects of EP and EE on the relative expression of jejunal proteins in broilers after immunization: a), b), c) is Western blot detects protein expression bands on days 7,21,35; d), e), f), g), h) is the level of the protein expression: TLR4,MyD88,TPAF6,ERK,JNK. All data were presented as mean ± SD(n = 6). *p < 0.05, **p < 0.01,***p < 0,001 compared to the group C; #p < 0.05,##p < 0.01,###p < 0,001 compared to the group V and intergroup.

Article Snippet: Name Company Cat. no. Dilution TLR4 Rabbit pAb Bioss bs-20379R 1:1000 MyD88 Rabbit pAb Novus Biologicals NB100-56698 1:1000 ERK1/2Rabbit pAb Abmart T55487 1:1000 JNK1/2/3Rabbit pAb Abmart T40073 1:1000 TRAF6Rabbit pAb ABclonal A16991 1:1000 GAPDHRabbit pAb ABclonal AC001 1:9000 Goat anti-Rabbit IgG Zenbio H&L 1:5000 X. Wang et al. Journal of Ethnopharmacology 319 (2024) 117306 On day 7 after vaccination, the FCR of the VE and VEE groups was significantly lower compared to the groups C and V (P < 0.05).

Techniques: Expressing, Western Blot